Method on how to perform pour plate method (no need dilution)

1. Collect one bottle of sterile molten agar (containing 15 mL of melted Plate Count Agar or any
other standard culture media) from the water bath (45°C).
2. Hold the bottle in the right hand; remove the cap with the little finger of the left hand.
3. Flame the neck of the bottle.
4. Lift the lid of the Petri dish slightly with the left hand and pour the sterile molten agar
into the Petri dish and replace the lid.
5. Flame the neck of the bottle and replace the cap.
6. Gently rotate the dish to mix the culture and the medium thoroughly and to ensure that the
medium covers the plate evenly. Do not slip the agar over the edge of the petri dish.
7. Allow the agar to completely gel without disturbing it, it will take approximately 10 minutes.
8. Seal and incubate the plate in an inverted position at 37°C for 24-48 hours.

Method on how to perform spread plate method (no need dilution)

1. Dip the L-shaped glass spreader into alcohol.

2. Flame the glass spreader (hockey stick) over a Bunsen burner.
3. Spread the sample evenly over the surface of agar using the sterile glass spreader, carefully
rotating the Petri dish underneath at the same time.
4. Incubate the plate at 37°C for 24 hours.
5. Calculate the CFU value of the sample. Once you count the colonies, multiply by the
appropriate dilution factor to determine the number of CFU/mL in the original sample.

3. To isolate single colony from microbial culture using quadrant streaking techniques

STREAKING
1. Take the petri dish. Open it, pour the agar in it around 3/4 of the petri dish. (make sure
sterillize the agar bottle's mouth through the flame first.)
2. Swirl it gently to make sure the agar is covering all of the petri dish.
3. Next take the inoculation loop and run it through the flame first. Then, dip it into your
sample.
4. Do the quadrant technique.